supporting information via rational designed upconversion ... · in a typical experiment,5 ml of...
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Supporting Information
Simultaneously High Sensitive Fluoride Ions Detection and Fluorosis Therapy
via Rational Designed Upconversion Nanoprobe
Yuxin Liu, Anqi Jiang, Qi Jia, Xuejiao Zhai, Lidong Liu, Liyi Ma, Jing Zhou*
Department of Chemistry, Capital Normal University, Beijing 100048, China
E-mail: [email protected]; Tel: +86-010-68902491
Table of Content
The supporting information contains a description of the further characterization and
condition investigation data in support of the manuscript.
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Figure S1. Size distribution of NaLuF4:Yb,Tm.
Figure S2. Size distribution of UCNP.
Figure S3. UCL spectra of NaLuF4:Yb,Tm and UCNP.
Figure S4. High-resolution TEM (HR-TEM) image A) and SAED patterns B) of
UCNP.
Figure S5. FTIR spectra of UCNP and UCNP-PA-Ti.
Figure S6. F and Ti amount in the supernatant of UCNP-PA-Ti-contained serum
within 90 days standing.
Figure S7. UCL spectra of UCNP-PA-Ti after receiving 10, 30, and 60 min
Electronic Supplementary Material (ESI) for Chemical Science.This journal is © The Royal Society of Chemistry 2018
irradiation.
Figure S8. I(470)/I(800) A) and I(470)/I(650) B) of pretreated UCNP-PA-Ti solution
with various concentration F- (medium concentration: 10 μM; high concentration: 20
μM) in BSA solution, HSA solution, serum, and urine.
Figure S9. I(470)/I(800) of UCNP-PA-Ti with cyanide and F- addition.
Figure S10. I(470)/I(800) of UCNP-PA-Ti in various basic pH conditions.
Figure S11. I(470)/I(800) and I(470)/I(650) of pretreated UCNP-PA-Ti solution with
F- in aqueous condition containing various biomolecules and anions.
Figure S12. I(470)/I(800) and I(470)/I(650) of UCNP-PA-Ti solution with various
soluble fluorides.
Figure S13. [TiF6]2-/total F- ratio of albumin-conjugated F- solution treated with
UCNP-PA-Ti.
Figure S14. Hydrodiameter of produced [TiF6]2-, CaF2, and MgF2 in vitro.
Figure S15. MTT assay and serum biochemistry tests.
Figure S16. H&E stained tissue sections.
Table S1. Hydrodiameter of UCNP-PA-Ti in various biological fluids and pH
conditions.
Table S2. F- capture efficiency of UCNP-PA-Ti in various anions presence.
Table S3. Comparison of the UCNP-PA-Ti and previous reported nanoprobes for F-
detection.
Table S4. UCL intensity of tubes presented in Figure 3E and F.
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1. Experiment section
1.1. Materials
Rare-earth oxide Lu2O3 (99.999%), Yb2O3 (99.99%), and Tm2O3 (99.99%) were
purchased from STREM Chemicals, Inc. USA. Na2SO4, KCl, MgSO4, CaCl2, FeSO4,
ZnCl2, NaCl, NaNO3, KBr, KI, Ti(SO4)2, NaOH, ethanol, cyclohexane, and
dichloromethane (CH2Cl2) were purchased from Beijing Chemical Reagent Company.
Oleic acid (OA), NaF, KF, nitrosonium tetrafluoroborate (NOBF4), glycine (Gly),
cysteine (Cys), lysine (Lys), serine (Ser), aspartic acid (Asp), dopamine hydrocloride
(DA), glucose (Glu), reduced glutathione (GSH), methyl cyanide, and ascorbic acid
(AA) were purchased from Sigma Aldrich. 1-Octadecene (ODE) and NH4F were
purchased from Alfa Aesar Chemical Co. Ltd. Bovine serum albumin (BSA), human
serum albumin (HSA), and pyrogallic acid (PA) were purchased from Energy
Chemical Co. Ltd. Rare earth chlorides (LnCl3, Ln: Lu, Yb, Tm) were prepared by
dissolving the corresponding metal oxide in HCl solution at elevated temperature and
then evaporating the water completely under reduced pressure. All other chemical
reagents were of analytical grade and were used directly without further purification.
Deionized (DI) water was used throughout.
1.2. Synthesis of NaLuF4:Yb,Tm@NaLuF4 nanoparticles (UCNP)
In a typical experiment, a mixture of 1 mM LnCl3 (Ln: 83%Lu, 16.9%Yb, and 0.1%
Tm), 15 mL OA, and 15 mL ODE were added into a 100 mL three-necked flask.
Under the vacuum, the mixture was heated to 160 oC to form a clear solution, and
then cooled to room temperature. After the solution cooling down, 0.025 mmol NaOH
and 0.04 mmol NH4F were added into the flask directly and stirred for 30 min. The
solution was slowly heated with gently stirred, degassed at 100 oC, and then heated to
300 oC and maintained for 1 h under the argon atmosphere. After the solution was
cooled naturally, the NaLuF4:Yb,Tm nanoparticles were separated via centrifugation
(10000 rpm) and washed with ethanol/cyclohexane (1:1 v/v) three times. The
hydrophobic NaLuF4:Yb,Tm was stored under room temperature in cyclohexane. The
UCNP with core-shell structure were obtained with the same method.
1.3. Synthesis of UCNP-PA-Ti
In a typical experiment, 5 mL of dichloromethane solution of NOBF4 (0.01 M) was
dropped into 5 mL of hydrophobic UCNP dispersion in cyclohexane (~5 mg mL-1) at
room temperature. Then, the bare UCNP were washed with ethanol several times and
dispersed in DI water. PA-Ti complex was obtained via a simple complexation
reaction by mixing 5 mL 5 mg mL-1 Ti(SO4)2 with 5 mL 10 mg mL-1 PA at room
temperature and ultrasonically treating for 5 min. Bare UCNP were then dispersed in
the solution of PA-Ti (pH = 7.4), being vigorous stirring for another 20 min at room
temperature. The resultant mixture was separated to obtain UCNP-PA-Ti via
centrifugation and washed with DI water several times to remove the extra PA-Ti
complex. The UCNP-PA-Ti was stored under 4 oC in DI water.
1.4. Characterization
The sizes and morphologies of NaLuF4:Yb,Tm and UCNP were determined using a
Tecnai G2F30 high-resolution transmission electron microscope (HR-TEM). Samples
of the UCNP were dispersed in cyclohexane and dropped on the surface of a copper
grid. The size distribution was counted and calculated from TEM images (α=0.90, 500
particles were measured) UV-vis-NIR absorption spectra were obtained on a
Shimadzu UV-3600 UV-vis-NIR spectrophotometer. Powder X-ray diffraction (XRD)
pattern was measured with a Brucker D8 advance X-ray diffractometer from 10o to
70o (Cu Kα radiation, λ = 1.54 Å). Dynamic light scattering (DLS) and zeta potential
experiments were carried out on an ALV-5000 spectrometer goniometry equipped
with an ALV/LSE-5004 light scattering electronic and multiple tau digital correlator
and a JDS Uniphase He–Ne laser (632.8 nm) with an output power of 22 mW. The
hydrodiameter distribution was measured at 25 oC with a detection angle of 90o. The
upcoversion luminescence (UCL) spectra were taken on a Maya LIFS-980
fluorescence spectrometer (Shanghai Oceanhood Opto-electronics tech Co. LTD)
equipped with an external 0-8 W 980 nm adjustable laser as the excitation source.
Inductively coupled mass spectroscopy (ICP-MS) analysis was performed on Agilent
7500ce ICP-MS.
1.5. Cell culture
CCC-HEL-1 cells were grown in DMEM (Dulbecco’s modified Eagle’s medium)
supplemented with 10% FBS (fetal bovine serum) and 1% penicillin-streptomycin at
37 oC with 5% CO2. For use in the MTT and oxidative stress experiments, 1 × 105
cells well-1 were seeded in 96-well plates and allowed to attach for 24 h prior to the
assay. For use in the in vitro experiments, 1 × 107 cells well-1 were seeded in 6-well
plates and allowed to attach for 24 h prior to the assay.
1.6. In vitro UCL imaging
CCC-HEL-1 cells with F- pretreated were incubated with UCNP-PA-Ti solution
(500 μg mL−1) for 5 h, washed with PBS several times, and were resupplied with fresh
DMEM. The UCL imaging was performed by optical microscopic study excited by
CW 980 nm. Signals were collected at 400-600 nm and 650 ± 15 nm, respectively.
1.7. In vivo UCL imaging
In vivo UCL imaging was performed with a modified in vivo luminescence imaging
system using an external 0–8 W adjustable CW infrared laser (980 nm, Hide-Wave
Co., China) as the excited source. Signals were collected at 400-600 nm and 800 ± 12
nm by Andor iXon Ultra EMCCD, respectively. Images of signals were analyzed with
corresponding software. Nude mice (m = 19 ± 2 g) were anesthetized with 10%
chloral hydrate (150 μL) and were intravenously injected with UCNP-PA-Ti solution
(10 mg kg-1). Undoped NaLuF4@NaLuF4-PA-Ti was used for regular administration.
At the 4 h post-injection, whole-body imaging was performed.
1.8. MTT assay
In vitro cytotoxicity was measured by performing methyl thiazolyl tetrazolium
(MTT) assays on the CCC-HEL-1 cells. Different concentrations of UCNP-PA-Ti (0,
2, 4, 6, 8 and 10 mg mL-1) were then added to the wells. The cells were subsequently
incubated for 24 or 48 h at 37 oC under 5% CO2. Thereafter, MTT (10 μL, 5 mg mL-1)
was added to each well and the plate was incubated for an additional 4 h at 37 oC
under 5% CO2. The optical density value (Abs.) of each well, with background
subtraction at 690 nm, was measured by means of a Tecan Infinite M200
monochromator-based multifunction microplate reader. The following formula was
used to calculate the inhibition of cell growth.
Cell viability (%) = (mean of Abs. value of treatment group/mean of Abs. value of
control)×100%
1.9. Hematology studies
All the animal procedures were in agreement with institutional animal use and care
committee and carried out ethically and humanely. The blood were harvested from
mice intravenously injected with UCNP-PA-Ti (n = 3, dose = 20 mg kg-1, Test) and
from mice receiving no injection for 1 day, 7 days, and 30 days post-injection (n = 3,
dose = 0 mg kg-1, Control), respectively. Blood was collected from the orbital sinus
by quickly removing the eye ball from the socket with a pair of tissue forceps. Five
important hepatic indicators (ALT, alanine aminotransferase; AST, aspartate amino
transferase; TBIL, total bilirubin; ALB, albumin; TP, Total protein) and one indicator
for kidney functions (CREA, Creatinine) were measured. Blood smears were prepared
by placing a drop of blood on one end of a slide, and using another slide to disperse
the blood along the length of the slide. Upon completion of the blood collection, mice
were sacrificed.
1.10. Hematoxylin and eosin (H&E) stained tissues
The heart, liver, spleen, lung, and kidney were harvested from mice intravenously
injected with UCNP-PA-Ti (n = 3, dose= 20 mg kg-1, Test) and from mice receiving
no injection (n = 3, dose = 0 mg kg-1, Control), 1 day, 7 days, and 30 days post-
injection, respectively. The tissues were fixed in paraformaldehyde, embedded in
paraffin, sectioned, and stained with H&E. The histological sections were observed
under an optical microscope.
Figure S1. Size distribution of NaLuF4:Yb,Tm.
Figure S2. Size distribution of UCNP.
Figure S3. UCL spectra of NaLuF4:Yb,Tm and UCNP.
Figure S4. High-resolution TEM (HR-TEM) image A) and SAED patterns B) of
UCNP. The SAED patterns were obtained from HR-TEM image via FFT.
Figure S5. FTIR spectra of UCNP and UCNP-PA-Ti. The band of UCNP at 1637 cm-
1 corresponds to stretching vibration of carbonyl. The bands of UCNP at 2851 cm-1
and 2922 cm-1 correspond to stretching vibration of C-H. New bands of UCNP-PA-Ti
at 1541 cm-1, 1612 cm-1, and 1634 cm-1 correspond to stretching vibration of C=C in
phenyl. The band of UCNP-PA-Ti at 3283 cm-1 corresponds to stretching vibration of
O-H.
Figure S6. F A) and Ti B) amount in the supernatant of UCNP-PA-Ti-contained
serum (1), 0.9% NaCl solution (2), 5% glucose solution (3), and artificial
cerebrospinal fluid (4) within 90 days standing. F C) and Ti D) amount in the
supernatant of UCNP-PA-Ti-contained PBS solutions with various pH within 90 days
standing.
Figure S7. UCL spectra of UCNP-PA-Ti after receiving 10, 30, and 60 min
irradiation.
Figure S8. I(470 nm)/I(800 nm) A) and I(470 nm)/I(650 nm) B) of pretreated UCNP-
PA-Ti solution with various concentration F- (medium concentration: 10 μM; high
concentration: 20 μM) in BSA solution, HSA solution, serum, and urine.
Figure S9. I(470)/I(800) of UCNP-PA-Ti with cyanide (UCNP-PA-Ti + cyanide) and
F- (UCNP-PA-Ti + F-) addition, respectively.
Figure S10. I(470)/I(800) of UCNP-PA-Ti in various basic conditions (pH = 7.0, 7.5,
8.0, 8.5, 9.0, and 9.5), respectively.
Figure S11. I(470 nm)/I(800 nm) A) and I(470 nm)/I(650 nm) B) of pretreated
UCNP-PA-Ti solution with F- in aqueous condition containing various biomolecules.
I(470 nm)/I(800 nm) C) and I(470 nm)/I(650 nm) D) of pretreated UCNP-PA-Ti
solution with F- in aqueous condition containing various anions.
Figure S12. I(470 nm)/I(800 nm) A) and I(470 nm)/I(650 nm) B) of UCNP-PA-Ti
solution with various soluble fluorides.
Figure S13. [TiF6]2-/total F- ratio of albumin-conjugated F- solution treated with
UCNP-PA-Ti within 4 min.
Figure S14. Hydrodiameter of produced [TiF6]2-, CaF2, and MgF2 in vitro.
Figure S15. A) Cytotoxicity of UCNP-PA-Ti determined by MTT assay. B-D) Serum
biochemistry tests including indices of liver and kidney function. Alanine
aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL),
creatinine (CRE), albumin (ALB), and total protein (TP) were tested.
Figure S16. H&E-stained tissue sections of major organs, including heart, liver,
spleen, lung, and kidney. Scale bar: 50 μm.
Table S1. Hydrodiameter of UCNP-PA-Ti in various biological fluids, including (1)
serum, (2) 0.9% NaCl solution, (3) 5% glucose solution, and (4) artificial
cerebrospinal fluid, and pH conditions (pH = 5.0 - 9.0).
H2O (1) (2) (3) (4)
HD (nm) 28.0 27.6 28.2 28.4 27.9
RSD (%) 1.346 1.667 2.614 2.639 1.271
5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5
HD (nm) 27.3 28.3 28.9 27.5 27.9 28.9 29.0 27.5 28.6 27.0
RSD (%) 1.789 2.674 1.528 1.468 2.389 2.128 2.400 1.349 1.863 2.505
Table S2. F- capture efficiency of UCNP-PA-Ti in various anions presence.
Cl- Br- I- SO42- CO32- NO3-
F- CaptureEfficiency (%) 84.3 ± 1.7 83.8 ± 2.2 85.4 ± 0.9 84.7 ± 2.3 83.9 ± 3.4 84.0 ± 2.7
Table S3. Comparison of the UCNP-PA-Ti and previous reported nanoprobes for F-
detection.
Strategy LOD (M) Linear Range (M) Reference
UCNP-PA-Ti ratiometric 4.2×10-9 5.0×10-8 – 2.6×10-5 this work
Eu-MOF ratiometric 2×10-6 4×10-6 – 8×10-5 [1]
CdS QDs-Ca turn-off 6×10-6 1×10-5 – 3×10-4 [2]
Eu(L–S15)2TTA turn-off 1.6×10-9 1.0×10-7 – 1.0×10-6 [3]
Table S4. UCL intensity of tubes presented in Figure 3E and F.
F- free 5 μM 10 μM 20 μM
I(470 nm) 3970.63 5786.52 6898.03 9083.40
I(650 nm) 4308.30 4619.54 4504.47 4417.51
I(800 nm) 15796.50 15837.48 15821.63 15132.10
I(470 nm)/I(650 nm) 0.9216 1.2526 1.5314 2.0562
I(470 nm)/I(800 nm) 0.2514 0.3654 0.4360 0.6003